search
for
 About Bioline  All Journals  Testimonials  Membership  News  Donations


Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 28, No. 5, 2007, pp. 470-476
Bioline Code: zr07072
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 28, No. 5, 2007, pp. 470-476

 en Effect of DNA Methylation on Igf-2r Expression in the Development of Cloned Cattle
CAI, Xia & LONG, Jian-er

Abstract

Epigenetic reprogramming has a crucial role in establishing nuclear totipotency in normal development and in cloned animals. In the current study, the method of real-time fluorescent quantitative PCR (FQ-PCR) was applied to detect Igf2r mRNA in Madin–Darby bovine kidney(MDBK )cells after being treated with 5'-azacytidine (5'-aza, a DNA methyltransferase inhibitor). And then we used the method of Bisulfite DNA Sequencing to detect DNA methylation status of Igf-2r DMR 2(DNA differentially methylated region,DMR)and 3'UTR(3'-untranslated region, UTR)in several tissues, such as brain, liver, lung, and heart in normal and cloned cattle.The results suggested that Igf2r mRNA were up-regulated in MDBK cells after being treated with 5'-aza. DNA methylation at DMR 2 significantly varied in normal cattle tissues but without significant variation at Igf-2r 3'-UTR. DNA methylation at Igf-2r DMR2 in cloned cattle was markedly altered compared with a normal fetus, while it was similar to a normal fetus at Igf-2r 3'-UTR. Based on these results, it is suggested that DNA methylation inhibitor down-regulated the expression of Igf-2r in MDBK cells. In normal cattle, patterns of DNA methylation at Igf-2r DMR2 was variable in different tissues, suggesting that the mechanism of gene imprinting was differently regulated in these tissues. The results also showed that in cloned cattle, Igf-2r DMR2 DNA methylation was disrupted while the non-imprinting control region (3'-UTR) was not. It suggested that disruption of the gene imprinting control region was likely to result in the abnormalities of cloned cattle.

Keywords
Igf-2r; Epigenetics; Cloned cattle; DNA methylation; Gene imprinting

 
© Copyright 2007 Kunming Institute of Zoology, the Chinese Academy of Sciences
Alternative site location: http://www.zoores.ac.cn/

Home Faq Resources Email Bioline
© Bioline International, 1989 - 2017, Site last up-dated on 05-Dec-2017.
Site created and maintained by the Reference Center on Environmental Information, CRIA, Brazil
System hosted by the Internet Data Center of Rede Nacional de Ensino e Pesquisa, RNP, Brazil